本文選題：兔大腸桿菌病 + 實質(zhì)器官�。� 參考：《山東農(nóng)業(yè)大學》2015年碩士論文

【摘要】：大腸桿菌病是由一定血清型的致病性大腸桿菌及其毒素引發(fā)的一種腸道傳染病。該病一年四季都能發(fā)生,感染各個年齡段的兔子,但主要對斷乳至4月齡仔兔的威脅最大。為進一步探討大腸桿菌對病兔實質(zhì)器官的影響,本實驗以人工感染的大腸桿菌仔兔為實驗對象,通過對患病仔兔實質(zhì)器官氧化損傷指標的檢測及免疫組化法(ICH)檢測不同發(fā)病時期病兔脾臟中CD14的分布情況,從一個新的角度研究致病性大腸桿菌對機體的病理損傷情況,為進一步防治大腸桿菌病提供新的臨床依據(jù)。本實驗將32只30~40日齡的新西蘭兔平均分為對照組與發(fā)病組兩組,發(fā)病組人工腹腔注射大腸桿菌原液,對照組腹腔注射等量生理鹽水,分別采取兩組第1、3、5、7d仔兔各實質(zhì)器官(心、肝、脾、肺、腎)1 cm3液氮速凍,保存于-80℃冰箱備用,用于檢測各實質(zhì)器官氧化損傷指標。分別測定各器官的SOD活性,MDA及蛋白含量,分析比較發(fā)病組與對照組各指標變化以及發(fā)病組不同發(fā)病時期各指標的變化規(guī)律。另取1cm3脾臟,用生理鹽水將實質(zhì)器官表面沖干凈后固定于40 g/L多聚甲醛中,制作石蠟切片,用于免疫組化法檢測仔兔脾臟中CD14的分布情況,統(tǒng)計仔兔脾臟的血管,白髓,紅髓,小梁CD14的陽性率,分析比較發(fā)病組與對照組CD14分布情況以及發(fā)病組不同發(fā)病時期陽性率的變化規(guī)律。病理解剖結(jié)果:病兔腹腔內(nèi)存在大量纖維蛋白,有明顯的臭味,腸壁外附著有大量的纖維蛋白,結(jié)腸壁有出血現(xiàn)象,腸內(nèi)糞便呈膠凍狀,肝臟和肺臟有明顯的充血、出血現(xiàn)象,脾臟外附著有大量纖維蛋白,個別病兔腎臟出現(xiàn)充血現(xiàn)象。氧化損傷指標結(jié)果:發(fā)病兔實質(zhì)器官SOD活性比對照組低,并且發(fā)病組隨病情發(fā)展呈先降低后升高的趨勢。肝臟在第一天SOD活性達到最低,心臟和脾臟在第三天達到最低,肺臟和腎臟在第五天達到最低。其中心臟在第三天與對照組存在極顯著差異(P0.01),肝臟在第一天與對照組相比存在顯著差異(P0.05),脾臟在第三天與對照組存在顯著差異(P0.05),肺臟和腎臟在第五天與對照組相比存在顯著差異(P0.05);蛋白含量發(fā)病組比對照組升高,發(fā)病組隨病情發(fā)展呈先升高后降低趨勢,其中心臟在第三天與對照組存在顯著差異(P0.05),肝臟在第一天與對照組存在顯著差異(P0.05),腎臟在第五天與對照組存在顯著差異(P0.05);MDA值比對照組升高,發(fā)病組隨病情發(fā)展呈先升高后降低的趨勢。肝臟在第一天達到最高,心臟和脾臟在第三天達到最高,肺臟和腎臟在第五天達到最高。其中心臟和脾臟在第三天與對照組存在顯著差異(P0.05),肺臟在第五天與對照組存在顯著差異(P0.05),變化趨勢與臨床表現(xiàn)相吻合。免疫組化結(jié)果:所采取的對照組脾臟有4組CD14檢測為陰性,其余12組均出現(xiàn)陽性;發(fā)病兔的脾臟中均檢測為陽性,并且陽性結(jié)果非常明顯,其中血管周圍分布最多,小梁及白髓分布較多,紅髓中分布較少。對脾臟中各部位CD14的陽性表達數(shù)據(jù)進行分析顯示,發(fā)病組較對照組表達量升高,并隨著病情發(fā)展,陽性表達率先升高后降低,在第三天和第五天的表達較高。其中白髓中的表達量在第三天比對照組顯著增加(P0.05);紅髓在第五天比對照組顯著增加(P0.05),第三天與對照組相比存在極顯著差異(P0.01);血管和小梁在第一天、第三天和第五天與對照組相比存在極顯著差異(P0.01)。實驗結(jié)果表明,健康仔兔人工感染大腸桿菌后,各實質(zhì)器官的氧化損傷指標均受到影響,并且不同器官的最高氧化損傷程度發(fā)生在不同發(fā)病時期。肝臟在第一天、心臟和脾臟在第三天、肺臟和腎臟在第五天的氧化損傷最高。免疫組化結(jié)果表明,仔兔在發(fā)病期脾臟中CD14敏感性增強,并在第三天達到最高,與氧化損傷結(jié)果相吻合,表明仔兔大腸桿菌除了對腸道產(chǎn)生損傷外,對實質(zhì)器官也產(chǎn)生了一定程度的損傷。
[Abstract]:Colibacillosis is an enteric infectious disease caused by a certain serotype of pathogenic Escherichia coli and its toxin. The disease can occur all the year round and infect rabbits of all ages, but it is the main threat to weaning to 4 month old rabbits. By detecting the oxidative damage index of the parenchyma organs of the infected rabbits and detecting the distribution of CD14 in the spleen of the sick rabbits at different onset period, the distribution of the infected rabbit's spleen was detected by the Immunochemistry method (ICH), and the damage of the pathogenic Escherichia coli to the body was studied from a new angle, so as to further prevent the colibacillosis. 32 30~40 day old New Zealand rabbits were divided into two groups in the control group and the onset group. The disease group was injected with the abdominal injection of Escherichia coli, and the control group was injected with equal amount of saline in the abdominal cavity. The 1 cm3 liquid nitrogen freezer of the two groups of 1,3,5,7d rabbits (heart, liver, spleen, lung and kidney) was taken respectively, and stored at -80 C ice. The SOD activity, MDA and protein content of each organ were measured respectively, and the changes of the indexes of the disease group and the control group and the changes of the indexes in the different onset period of the disease group were analyzed and compared. The spleen of the 1cm3 and the surface of the parenchyma organs were washed and fixed to 40 g/L after the surface of the parenchyma organs were washed with saline. Paraffin section was made to make paraffin slices, and the distribution of CD14 in the spleen of rabbit was detected by immunohistochemistry. The positive rates of blood vessels, white pulp, red pulp and trabecular CD14 in rabbit spleen were statistically analyzed. The distribution of CD14 and the change of positive rate in different onset groups of the disease group and the control group were analyzed and compared. The pathological anatomical results: the abdominal cavity of the sick rabbit There is a large amount of fibrin in a large number of fibrin, a large number of fibrin protein attached to the wall of the intestines, bleeding in the colon wall, gelatinous feces in the intestines, obvious congestion in the liver and lungs, bleeding, a large amount of fibrin attached to the spleen outside the spleen, and the phenomenon of hyperemia in the kidney of a few sick rabbits. The result of the oxidative damage index: the rabbit parenchyma The SOD activity of the organs was lower than the control group, and the onset group decreased with the development of the disease first and then increased. The liver had the lowest SOD activity on the first day, the heart and spleen reached the lowest in third days, the lungs and the kidneys reached the lowest in the first day. The heart was significantly different from the control group on the third day (P0.01), the liver was on the first day. There was significant difference in the control group (P0.05), the spleen was significantly different from the control group at third days (P0.05), the lungs and kidneys were significantly different from the control group on the fifth day (P0.05); the protein content group was higher than the control group, and the onset group increased with the development of the disease, and the heart was stored at third days with the control group. In the significant difference (P0.05), the liver was significantly different from the control group on the first day (P0.05), the kidney was significantly different from the control group at fifth days (P0.05); the MDA value was higher than the control group. The incidence of the disease was higher and then decreased with the development of the disease. The liver reached the highest on the first day, the heart and spleen reached the highest in the third day, lung and kidney. There was a significant difference between the fifth days in the fifth days. The heart and spleen were significantly different from the control group on the third day (P0.05), and there was a significant difference between the control group and the control group on the fifth day (P0.05). The change trend was consistent with the clinical manifestation. The immunization results showed that there were 4 groups of CD14 in the spleen of the control group and the other 12 groups were positive. The positive results of the rabbit's spleen were positive, and the positive results were very obvious. The distribution of the trabeculae and the white pulp were most, the distribution of the trabeculae and the white pulp were more, the distribution of the positive expression of CD14 in the spleen was higher than that of the control group, and the positive expression was first increased with the development of the disease. The expression in the third and fifth days was higher in the third and fifth days than in the control group (P0.05), the red pulp increased significantly on fifth days (P0.05), and the third day was significantly different from the control group (P0.01); the blood vessels and trabeculae were compared with the control group at the first, third and fifth days. The experimental results showed that after artificial infection of Escherichia coli in healthy rabbits, the oxidative damage indexes of all the parenchymal organs were affected, and the highest degree of oxidative damage in different organs occurred at different times. On the first day of the liver, the heart and spleen were third days, the lungs and kidneys were the most oxidative damage in the fifth day. The results of immuno histochemistry showed that the sensitivity of CD14 in the spleen increased at the onset of the onset and reached the highest in third days, which was consistent with the results of oxidative damage. It showed that the Escherichia coli had a certain degree of damage to the parenchyma in addition to the damage to the intestine.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.291

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本文編號：2100220